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  • Title: [Study on culture, identification and differentiation of CD133+ endothelial progenitor cells from human umbilical cord blood].
    Author: Huang Y, Wang SM, Wang JS, Huang XL.
    Journal: Zhonghua Wai Ke Za Zhi; 2007 May 01; 45(9):619-22. PubMed ID: 17688798.
    Abstract:
    OBJECTIVE: To study the isolation, culture and identification of CD133+ endothelial progenitor cells (EPCs) from human umbilical cord blood in vitro. METHODS: EPC separation was performed with density gradient centrifugation and MACS separation. Purity of EPCs was determined by flow cytometry. EPC was cultured with EBM-2 to study the cultivate features of EPC. Uptake test of Dil-LDL and FITC-Lectin and immunohistochemistry were performed. RESULTS: According to flow cytometry, (1.13 +/- 0.10)% of mono-nuclear cells were CD133+ and the purities of CD133+ EPCs were (91.45 +/- 1.04)% on average. CD133+ EPCs became adherent, spindle-shaped and formed cluster during culture. Uptake test of Dil-LDL and FITC-Lectin were positive. (95.83 +/- 1.72)% of CD133+ cells were found positive in both uptake tests. The positive rates of immunostaining of cell markers CD34 and factor VIII were (95.83 +/- 2.23)% and (95.92 +/- 1.43)% after cultured for one week, which showed no significant differences between CD133+ EPCs and human umbilical vein endothelial cells. Capillary structures were formed by CD133+ EPCs after cultured for 4 and 7 d in vitro. CONCLUSIONS: High purity of CD133+ EPCs can be obtained by MACS separation. CD133+ EPCs can differentiate into mature endothelial cells with the effects of stimulating factors.
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